title: Horse heart myoglobin reconstituted with a symmetrical heme. A circular dichroism study.
abstract: Proton NMR studies on myoglobins and hemoglobins reconstituted with non-natural hemes, possessing different side chains in the pyrrolic rings, have provided interesting information for the understanding of the mechanism governing heme reorientation in the globin pocket, during synthesis of the native protein in vivo or in the reconstitution process in vitro. More recently, circular dichroism (CD) studies have been reported as a qualitative, alternative tool, with respect to 1H-NMR for detecting heme disorder in a reconstituted myoglobin or hemoglobin. In this paper, a CD study is reported on the reconstitution of horse heart myoglobin with protoheme XIII, a heme possessing true rotational symmetry about its alpha, gamma-meso axis. The results obtained show that the reconstitution product with this heme, which binds to the apoprotein with high affinity, not dissimilar from that of the natural heme, is characterized by a CD spectrum with bands possessing rotational strengths much lower than in the native protein. Furthermore, the CD changes detected as a function of time, during heme reorientation, in the case of natural heme, are absent when the apoprotein is reconstituted with protoheme XIII. These data provide independent evidence for reorientation of the natural heme, which follows its insertion into the protein matrix.
authors.affiliation: Department of Biochemical Sciences, University La Sapienza, Rome, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Santucci
authors.forename: F
authors.initials: F
authors.lastname: Ascoli
authors.forename: G N
authors.initials: GN
authors.lastname: La Mar
authors.forename: D W
authors.initials: DW
authors.lastname: Parish
authors.forename: K M
authors.initials: KM
authors.lastname: Smith
mesh_terms: D000818:Animals; D001059:Apoproteins / Q000378:metabolism; D002942:Circular Dichroism; D006418:Heme / Q000378:metabolism*; D006736:Horses; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D009206:Myocardium / Q000378:metabolism; D009211:Myoglobin / Q000378:metabolism*; D011487:Protein Conformation; D013053:Spectrophotometry
chemical_list: D001059:Apoproteins; D009211:Myoglobin; C010857:apomyoglobin; D006418:Heme